GSM7384417: Brain_EV; Mus musculus; RNA-Seq
Tissue was digested in with Accutase (Merck) at 37 °C for 30 min and triturated through 70 μm cell strainers which were rinsed with 10% FCS in PBS. Cells were purified by a linear 40% Percoll (GE Healthcare) centrifugation step at 650 xg without brakes for 25 min and the supernatant were discarded. Mononuclear cells were resuspended in FACS buffer (1% BSA and 0.1% sodium azide in PBS) and stained for live cells, CD45, CD11c, CD8, and CD4. Cells from similar organs and treatment were pooled from five mice and single cell libraries prepared. Single cells were encapsulated in droplets using 10X Genomics GemCode Technology kit v2 and processed following manufacturer's specifications. Chromium single cell 3´ v2 (10XGenomics)